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Research progress in the application of deep learning methods for marine fishery production: a review
ZHANG Shengmao, SUN Yongwen, FAN Wei, TANG Fenghua, CUI Xuesen, WU Yumei
Journal of Dalian Ocean University    2022, 37 (4): 683-695.   DOI: 10.16535/j.cnki.dlhyxb.2022-099
Abstract （5534）   HTML （31）    PDF（pc） （3946KB）（3781）       Save
With the continuous decline of global fishery resources, fishery agencies and regional fisheries management organizations of various countries use the fishery observer method to promote sustainable fishing, but the human observer method is characterized by so high cost and low coverage as to difficult to meet the management needs. In recent years, detection speed and accuracy have been continuously enhanced due to continuous emergence and improvement of new deep learning algorithms, thus providing conditions for marine fishery fishing production monitoring. The process of building a fishery production monitoring model is introduced from the aspects of data acquisition, data preprocessing, algorithm design, model training, and model accuracy evaluation. The application of deep learning technology in marine fishery fishing is discussed, and methods such as transfer learning or reinforcement learning are proposed in terms of fishing boats and boat behavior, catches, fishery forecasts, crew members, and fishing gear to expand the identification of target species and enhance detection algorithm，and use high-accuracy feature extraction network to improve the accuracy of target classification, to solve the real-time analysis of electronic monitoring data through edge computing technology and to formulate standards and specifications for electronic monitoring fishery management applications and other key research directions in the future, which provides reference for the promotion of deep learning in marine fishery fishing production.
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Progress on research and control of amyloodiniosis in marine fish: a review
LI Zhicheng, ZHONG Zhihong, LI Shiyu, GUO Yixuan, GUO Qingkai, JIANG Biao, LI Anxing
Journal of Dalian Ocean University    2022, 37 (4): 696-706.   DOI: 10.16535/j.cnki.dlhyxb.2021-190
Abstract （5129）   HTML （10）    PDF（pc） （5437KB）（3781）       Save
Amyloodiniosis, commonly referred to as “marine velvet”, velvet disease, is a parasitological disease caused by one of the most common and important parasitic dinoflagellates  Amyloodinium   ocellatum . The  A.ocellatum  infects a wide range of fish species in different habitats including tropical and subtropical geographical locations, and has a direct life cycle that comprises three stages: trophont, tomont, and dinospores, which can be completed in less than a week dependent on favorable environmental factors. The disease is characterized by rapid onset and high mortality, which seriously hinders the healthy development of China’s marine fish industrial farming and semi-intensive pond farming, leading to huge economic losses. The typical symptoms of amyloodiniosis are changes in fish behavior, decreased appetite, faster breathing, and swimming at the water surface or gathering at the areas with higher dissolved oxygen concentrations. Fish mortalities are usually attributed to hypoxia, inflammation, severe gill hyperplasia, hemorrhage, and necrosis in heavy infestations. This review summarizes the current situation and the latest researches progress on the life cycle of  A.ocellatum , the prevalence, detection methods, and preventive and control measures of amyloodiniosis. It is proposed to strengthen the future research of pathogenic detection of the farming environment, low-temperature preservation, vaccine development, aiming to provide references for the scientific prevention and treatment of marine fish amyloodiniosis in fisheries production.
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Organisms attachment on 3D printed concrete artificial reefs
SHEN Lu, ZHANG Nianhua, TIAN Tao, HONG Hexuan
Journal of Dalian Ocean University    2022, 37 (4): 584-591.   DOI: 10.16535/j.cnki.dlhyxb.2022-073
Abstract （5067）   HTML （24）    PDF（pc） （2283KB）（3674）       Save
In order to study the organisms attachment of 3D printed concrete artificial reef, the species and biomass of attached organisms were comparatively investigated on 3D printed hanging plate specimens with nozzle diameters of 20 mm (3D-20 group) and 10 mm (3D-10 group) and cast-in-place hanging plate specimens with the same mix proportion (PC group)disposed in Heishijiao sea area of Dalian, Liaoning Province on November 1, 2020 (for 2 months), February 1, 2021 (for 3 months), and July 1, 2021(for 5 months). After the 1st and 2nd sampling, the specimens were put back in place. The offshore hanging board were completed on September 1, 2020. A total of 17 species of attached organisms were sampled and identified, with dominant species snail  Littorina   brevicula  in each sampling. The total biomass was shown to be 561.58 g/m 2  in 3D-20 group, 215.72 g/m 2  in 3D-10 group, and 203.24 g/m 2  in PC group in the third sampling. For three samplings, there was significantly higher total biomass in 3D-20 group than that in 3D-10 group and PC group, significant difference between 3D-20 group and 3D-10 group and PC group ( P <0.05), and no significant difference between 3D-10 group and PC group ( P >0.05). The finding indicates that 3D printed concrete is practical as artificial reef material, and the roughness of reef surface caused by printing method is superior to biological attachment.
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Research advancement on degradation mechanism and ecological restoration technology of coastal salt-marsh：a review
ZHANG Mingliang
Journal of Dalian Ocean University    2022, 37 (4): 539-549.   DOI: 10.16535/j.cnki.dlhyxb.2022-181
Abstract （5054）   HTML （53）    PDF（pc） （14224KB）（3650）       Save
Coastal salt-marsh is one of the important ecosystems connecting land and sea, and has significant ecological functions and ecological values. In recent years, a series of factors have contributed to the serious degradation in coastal salt-marsh. Exploration and practice on the degradation causes and restoration technologies in coastal salt-marsh have been conducted considerable studies from different fields, with a huge number of study results. Ecology, hydrology, environmental science, biology, estuarine dynamics, and other disciplines are all involved in the degradation and restoration technology of coastal salt marsh, being a typical interdisciplinary and hot research field. Here, we summarize the growth and development process of wetlands based on an analysis of the distribution characteristics of coastal salt marsh in China, and probe into the interaction between salt marsh plants and their influencing factors in terms of salinity stress, hydrological conditions, natural factors, natural succession, and human activities, which deepens the understanding of degradation factors and mechanisms in coastal salt marsh. We generalize the existing research on ecological restoration technology in coastal salt marsh, put forward the future prospects, such as wetland degradation mechanisms from a multi-disciplinary perspective, tridimensional and continuous studies using a variety of technologies, integrated application of numerous ecological restoration technologies, and discuss risk assessment of  Spartina   alterniflora  intrusion to provide a scientific reference for the research on degradation and ecological restoration in coastal salt marsh.
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Artificial propagation and embryonic and larval development of Chinese hook snout carp Opsariichthys bidens
LUO Xiaonian, DUAN Youjian, ZHANG Xiaojing, ALIDA·Xiamuxiding, LI Jiao, WU Chen, LI Wenkang, JI Chenyue, CHANG Jiantai
Journal of Dalian Ocean University    2022, 37 (4): 550-557.   DOI: 10.16535/j.cnki.dlhyxb.2021-222
Abstract （6294）      PDF（pc） （4024KB）（3630）       Save
In order to realize the whole process of artificial propagation and culture of wild Chinese hook snout carp  Opsariichthys   bidens , the fish was induced to spawn by hormone injection, the fertilized eggs were incubated and fry and juveniles were reared in a pond at Xingda Farm in Liaoyang County, Liaoning Province from 2018 to 2021.Meanwhile embryonic and larval development was observed under a microscope. It was found that male fish were maturity at 1 +  year old, and the female at 2 +  years old in a pond culture. The pond-cultured Chinese hook snout carp spawned from May 10th to May 29th in Liaoyang area, and had average success rate of 73.8%, fertilization rate of 81.3% and hatching rate of 78.8% by injection of mixed drugs (LHRH-A 2  4-8 μg/kg+DOM 4-8 mg/kg+HCG 1 000-2 000 IU/kg), artificial dry insemination, combined hatching basket and hatching bucket incubation method. The fertilized eggs of Chinese hook snout carp are orange-yellow in color without adhesion, and the spawners had effect time of 25 to 27 h after hormone injection at water temperature from 18 to 20 ℃ (one injection). The embryonic development was divided into 7 stages including blastoderm, cleavage, blastula, gastrula, neuroblastoma, organogenesis, and hatching and 24 sub-stages, lasting about 62 h 6 min, with total incubation accumulative temperature of 1 199.91 ℃·h. The larvae swam horizontally over 5 to 6 days post hatching, and the 1-day-old Chinese hook snout carp larvae had total length of (5.13±0.63)mm. The larvae became the mixed nutrition period at the 10-day-old age, and the 12-day-old postlarvae had exhaustion of the yolk sac, and allometric growth. Total 14 000 F1 fry were obtained in the ponds in 2018, and 2 120 000 F2 fry were reared for two consecutive years from 2020 to 2021. The findings realized the whole procedure of artificial propagation of Chinese hook snout carp for the first time.
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Preparation and properties of ratio fluorescent probe for polarity detecting of biological microenvironment
LI Miao, JIN Xiaoyi, JIANG Shuo, SUN Meng, TIAN Ye
Journal of Dalian Ocean University    2022, 37 (4): 602-610.   DOI: 10.16535/j.cnki.dlhyxb.2021-323
Abstract （4469）   HTML （45）    PDF（pc） （5272KB）（3629）       Save
 To construct a polar ratio fluorescent probe for accurate detection of biological microenvironment, the probe IA based on naphthalene ring fluorophore was firstly synthesized by introducing malononitrile and N, N-dimethyl group on acenaphthene ketone, and then the polarity responsive ratio fluorescent probe IAN was synthesized by condensation reaction of IA amino group into heterocycle. The molecular structure and mass of probes IA and IAN were characterized by  1 H NMR,  13 C NMR and HRMS, and then the absorption and fluorescence spectra of probes IA and IAN in different polarity solvents were examined and compared by UV-Vis and fluorescence spectrophotometer. The results showed that IA exhibited single absorption and fluorescence emission peaks and fluorescence intensity of IA was gradually enhanced with solvent polarity increasing. Differently, double absorption and fluorescence emission peaks of IAN were simultaneously detected in solvents ( λ em =474 nm, and 552 nm), and the fluorescence intensity ratio  I 474 nm / I 552 nm  produced a sensitive Boltzmann function response to solvent polarity changes. Compared with IA, IAN was able to respond accurately to environmental polarity changes as a ratio fluorescent probe. The fluorescence intensity of the lesions was shown to be significantly lower than those of the healthy tissues by using IAN on mice with breast cancer and grouper hybrid  Epinephelus   fuscoguttatus ♀× E.lanceolatus ♂ with hepatobiliary syndrome. These findings indicated that amino group undergoing a condensation reaction to generate a heterocycle containing nitrogen atom increased the conjugation degree of lone pair electrons of the nitrogen atom to the π system, so that the probe IAN produced n→π *  and π→π *  molecular transition at the same time, and then enabled the construction of a ratio fluorescent probe responding sensitively to polarity changes.
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Nematode infection status and temporal characteristic of estuarine tapertail anchovy Coilia nasus in Anqing section of the Yangtze River
DAI Pei, GUO Qi, TIAN Jiali, MA Fengjiao, WANG Yinping, LIU Kai
Journal of Dalian Ocean University    2022, 37 (3): 464-470.   DOI: 10.16535/j.cnki.dlhyxb.2021-147
Abstract （4583）   HTML （71）    PDF（pc） （3509KB）（3622）       Save
In order to grasp nematode infection status of estuarine tapertail anchovy  Coilia   nasus  in the Yangtze River,the nematode infection was surveyed in 1 658 samples of  C.nasus  collected at the Anqing section of the Yangtze River from April to July 2018 for 72 d.The results showed that the infection rate of the whole samples was 96.32%,the average infection intensity was (10.69±9.67)individuals per fish,the average infection abundance was (10.16±9.67) individuals per fish.In the infected  C.nasus ,52.53% of individuals were infected with 1-9 nematodes,and a few fish infected with nematodes more than 30,accounting for only 3.93%.Nematodes are parasitic on the body cavity and organs of  C.nasus ,including the stomach,pyloric caecum,and intestinal tract.The gonads and liver are attached to the surface or embedded in the tissue.There was the maximal nematodes in intestine and pyloric caecum,and the proportions nematodes were 41.26%,and 31.15% respectively.The results of nematode sampling and identification showed that  C.nasus  was infected with 6 species of 2 genera,including 2 species of  Anisakis  and 4 species of  Hysterothylacium ,the dominant species is  Anisakis   pegreffii .The infection intensity of  C.nasus  in different sizes was different.With increase in body length,the infection intensity of  C.nasus  showed a trend of increasing first and then decreasing.The infection rate was the highest in the body length group of 280-300 mm,and the average intensity and abundance were the highest in the 260-279 mm body length group.In addition,during the investigation period,as time passed,the infection index of  C.nasus  was first increased and then decreased,and the infection was the most serious in the first half of June.In summary,this study has basically grasped nematode infection status of  C.nasus  in the Yangtze River,and may have a positive impact on the follow-up study which focused on the impact of nematode parasitism,and the  C.nasus  population dynamics after infected with nematode.
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Effects of compound phosphorus-free water retention agent on water retention of unrinsed minced flesh of sea bass Lateolabrax japonicus
BAO Yirui, ZHAO Qiancheng, WU Yanyan, WANG Yueqi, MAI Zhicheng
Journal of Dalian Ocean University    2022, 37 (4): 676-682.   DOI: 10.16535/j.cnki.dlhyxb.2021-178
Abstract （4883）   HTML （45）    PDF（pc） （3269KB）（3612）       Save
 In order to improve the water-holding quality of minced flesh of sea bass  Lateolabrax   japonicus , the effects of NaHCO 3 , trehalose and soybean protein isolate on water retention were investigated in unrinsed sea bass minced flesh via cooking loss, texture profile analysis (TPA) and sensory evaluation in single factor test and  L 9 (3 4 ) positive test by cross-experiment, and the optimal formula of three phosphorus-free water retention agents was determined. The microstructure of minced flesh products was observed by electron microscope. The results showed that the appropriate addition of three phosphorus-free water-retaining agents, NaHCO 3 , trehalose and soybean protein isolate led to enhance the water-retention of minced flesh. Too large amount, however, resulted in a certain impact on the quality, leading to the decrease in each index. The orthogonal test showed that the optimal formula of the compound phosphorus-free water-retaining agent was composed of 0.1% sodium bicarbonate and 0.3% trehalose (all in terms of mass fraction). The sensory score of the minced flesh product prepared with 3.5% soybean protein isolate and the addition of the compound phosphorus-free water-retaining agent was 95.8, the cooking loss was reduced by 43.4% compared with the blank group, and the water-retention effect was significantly improved. The microstructure observation showed that relatively compact and the small cavity were observed in the tissue structure of the minced flesh supplemented with the compound phosphorus-free water-retaining agent. The findings indicated that addition of a compound phosphorus-free water-retaining agent to unrinsed sea bass minced flesh led to effectively improve its water retention.
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Screening, identification and culture medium optimization of a hypoglycemic Lysinibacillus fusiformis strain from common carp
YU Junfei, XIA Banghua, HAO Qirui, ZOU Haobo, WANG Peng, CHEN Zhongxiang, GAO Lei, WU Song, HUANG Li, QIN Dongli, HAN Ying
Journal of Dalian Ocean University    2022, 37 (4): 611-619.   DOI: 10.16535/j.cnki.dlhyxb.2021-281
Abstract （5290）   HTML （41）    PDF（pc） （5304KB）（3582）       Save
 In order to investigate the hypoglycemic function of bacillus  Lysinibacillus   fusiformis  and thus enhance its metabolite concentration, bacteria with proteolytic ability were screened from common carp  Cyprinus   carpio  intestinal contents by protease production ability assay, and their ability to produce dipeptidyl peptidase-Ⅳ (DDP-Ⅳ) inhibitor was measured. The screened strains were isolated, purified and cultured, then identified by 16S rRNA sequencing and physiological and biochemical characteristics, and the medium formulation of the strains was by single-factor test and response surface optimization experiment. The results showed that the CR-3 strain, with the ability to produce 62.35% at 37 ℃, was identified as  L.fusiformis  by physiological and biochemical identification and phylogenetic tree construction. The optimum fermentation medium was consisted of 1.46% sucrose, 0.55% CaCO 3 , 0.48% NH 4 Cl and 2.10% soybean meal, with 64% higher DDP-Ⅳ inhibitor yield than the strain unoptimized. The findings indicate that the  L.fusiformis  screened in this experiment has good ability to produce DDP-IV inhibitor and has potential hypoglycemic function.
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Effects of bleeding on quality enhancement for freshness-locked in rock greenling Hexagrammos otakii
LIU Yuepeng, LIU Junrong, CAI Yan, QI Lin, XU Tanye, TIAN Yuanyong, QU Qinkun
Journal of Dalian Ocean University    2022, 37 (4): 668-675.   DOI: 10.16535/j.cnki.dlhyxb.2021-234
Abstract （5290）   HTML （70）    PDF（pc） （8102KB）（3561）       Save
 In order to explore enhancement effect of bleeding treatment on freshness-locked effect, the healthy living rock greenling  Hexagrammos   otakii  were bleeding for 10 min by spinal cord cut and then kept in ice water cooling (well-bled, WB), and in conventional water cooling immediately after the spinal cord cut (poor-bled, PB) and the healthy living rock greenling was not bled as suffocation group (none-bled, NB). The two freshness locked groups were divided into two parts, one part was stored with ice for the observation of the characteristics of post mortem rigor mortis process, and the other part was placed in a 4 ℃ environment for cold storage stability analysis. The fish quality was evaluated by monitoring the biochemical indicators including contents of ATP and glycogen and rigor mortis process, combined with sensory analysis. Results showed that addition of room temperature water bleeding treatment before the fish body cooled led to significantly improve the bleeding effect: there was significantly higher average blood output of the enhanced freshness locked in group WB (8.6 g/ind.±3.1 g/ind.) than that in the conventional freshness-locked group PB (5.7 g/ind.±2.3 g/ind.)( P <0.05), enhanced bleeding showing superior cold storage stability. The various stages of the process of post-mortem rigor mortis were significantly delayed by enhanced bleeding effect can. During ice storage, the initial time of rigor mortis was about 4 hours later in the group WB than that in the group PB, and the time of rigor off was also about 6 hours later in the group WB. There was better preservation effect on ATP in the group WB, with ATP content of 1.20 μmol/g in the group WB, and only 0.58 μmol/g in the group PB 12 hours after chilled storage. Enhanced bleeding treatment resulted in significant improvement of the sensory quality of fish. During chilled storage, the abdominal muscles in the group WB showed bright silvery white in color, and the abdominal muscles in the group NB with none blood discharge had obvious extravasated blood, with mediate in the whiteness of the abdominal muscles in the group PB. In summary, the disposal of room temperature water can effectively promote the bleeding of the fish, and significant freshness-locked effect in terms of cold storage stability and sensory quality.
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Recognition algorithm of marine ship based on improved YOLOv5 deep learning
ZHANG Xiaopeng, XU Zhiyuan, QU Sheng, QIU Wenxuan, ZHAI Zeyu
Journal of Dalian Ocean University    2022, 37 (5): 866-872.   DOI: 10.16535/j.cnki.dlhyxb.2022-104
Abstract （5991）   HTML （199）    PDF（pc） （7354KB）（3524）       Save
In order to improve the accuracy of marine ship recognition in multiple targets and foggy environments, a marine ship recognition model SE-NMS-YOLOv5 is proposed based on improved YOLOv5 deep learning. The model is combined with Dark channel defogging algorithm, SE(squeeze-and-congestion) attention mechanism module and improved non-maximum suppression model for training and testing of ship data sets. The results showed that in the ship recognition task, there was the accuracy of 90.6%, recall rate of 89.9% and SE-NMS-YOLOv5  F 1 value of 90.5%, and compared with YOLOv5 model, the detection effect is improved by 6.3%, 4.8% and 5.8%. Compared with YOLOv4, the model improved 19.1%, 19.0% and 19.3%. In foggy conditions, the accuracy, recall rate and  F 1 value of SE-NMS-YOLOv5-Dark channel model were 88.1%, 87.2% and 87.6%, compared with SE-NMS-YOLOv5 model, the detection results are improved by 13.8%, 13.3% and 13.5%, respectively. The findings indicate that the marine ship recognition method based on SE-NMS-YOLOv5 effectively solves the problem of low accuracy of marine ship detection on multiple targets and foggy conditions, and improve the overall effect of ship detection and recognition.
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Effects of crude protein from mucus of the whelk Volutharpa ampullacea Perryi on proliferation of hepatocellular carcinoma and tumor growth of tumor-bearing mice
YU Linghui, ZHAO Tanjun, CHEN Simeng, LI Na, XU Jing, LI Chunmao, ZHAN Yaoyao, CHANG Yaqing
Journal of Dalian Ocean University    2022, 37 (4): 592-601.   DOI: 10.16535/j.cnki.dlhyxb.2021-284
Abstract （5180）   HTML （75）    PDF（pc） （10099KB）（3520）       Save
To preliminarily clarify the anti-tumor effects of crude protein from the mucus of the whelk  Volutharpa   ampullacea  Perryi, the effects of crude protein from the mucus of the whelk on the proliferation of human hepatoma Hep-G2 cell and tumor growth of H 22  tumor-bearing mice were analyzed and compared in vitro and in vivo experiments using microscopic observation, flow cytometry and qRT-PCR techniques. The results showed that the morphology of Hep-G2 cells in the group treated with 0.66 g/L crude protein from the mucus of the whelk showed severe deformation, nuclear condensation and blurring of nuclear membrane, vacuolation of cytoplasm, swelling of mitochondria and significant reduction of microvilli on the cell surface compared with the untreated control group (as control). The proportion of G2/M stage cells in Hep-G2 cells in the group treated with 0.66 g/L crude protein from the mucus of the whelk was significantly higher than that in the untreated group ( P <0.05). The transcript levels of cell cycle protein  CCNB 1 and cell cycle protein-dependent kinase  CDK 1 in the Hep-G2 cells were shown to be significantly lower in the crude protein-treated group than those in the control group ( P <0.05). The mean tumor weights of the tumor-bearing mice were significantly lower in the crude protein-injected groups (the injection dose of prevention group and the treatment group was 30 mg/kg) than those in the model control group ( P <0.01). The findings indicate that crude protein from the mucus of the whelk has anti-hepatocellular carcinoma effects both in vitro and in vivo, showing the potential to be developed as an anti-hepatocellular carcinoma drug of marine origin.
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Heavy metal contamination and health risk assessment of tiger puffer Takifugu rubripes farmed in a recirculating tank
WANG Xuankai, ZHAO Tong, YUE Yang, WANG Houyu, QU Baocheng, XIONG Yuyu, ZHANG Yufeng
Journal of Dalian Ocean University    2022, 37 (4): 650-657.   DOI: 10.16535/j.cnki.dlhyxb.2021-280
Abstract （6113）   HTML （44）    PDF（pc） （1717KB）（3514）       Save
In order to evaluate the quality and safety of tiger puffer  Takifugu   rubripes  farmed in a recirculating tank, the contents of lead (Pb), cadmium (Cd), mercury (Hg), copper (Cu) and chromium (Cr) were determined in tiger puffer with average body weight of 2.3 g (juvenile), 1 091.4 g(sub-adult), and 1 718.1 g (adult) sampled from an industrial aquaculture enterprise in Dalian, Liaoning Province at different developmental by inductively coupled plasma mass spectrometry (ICP-MS). The results showed that the contents of different heavy metals in the body of the tiger puffer were not shown to exceed the safety standards of “National standards for food safety, limits of pollutants in food”(GB 2762—2017) and “Limits of toxic and harmful substances in non polluted food and aquatic products”(NY 5073—2006). The changes in heavy metal contents in the tiger puffer in different culture stages showed that Cu and Cr were the main metal elements enriched in liver ( P <0.05) and Cr in muscle ( P <0.05), without significant difference in contents of Cd, Hg, Cu and Cr in tiger puffer in different developmental stages ( P >0.05), and with significant difference in the contents of Cd, Hg and Cu in different tissues ( P <0.05). The assessment of heavy metal pollution in fish body showed that single factor pollution index and Nemerow comprehensive pollution index were in the non-pollution range, and according to the estimated daily intake, carcinogenic risk and non-carcinogenic risk, the contribution of daily intakes to health risks is expected to be small, cancer risk in children and adults is within acceptable limits, and non-cancer risk is negligible, but the overall health risk in children is slightly higher than that in adults. The findings indicate that the levels of heavy metals in tiger puffer can be controlled within a safe range in the recirculating aquaculture system.
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Expression analysis and preparation of recombinant protein of T-cell immunoglobulin and mucin-domain-containing molecule 4 (TIM-4) gene from oblique grouper Epinephelus coioides
LI Jinze, TANG Jufen, HUANG Yu, JIAN Jichang, SHI Gang, CAI Jia
Journal of Dalian Ocean University    2022, 37 (3): 403-410.   DOI: 10.16535/j.cnki.dlhyxb.2021-172
Abstract （5790）   HTML （68）    PDF（pc） （7037KB）（3496）       Save
In order to study the role of T-cell immunoglobulin and mucin-domain-containing molecule 4 (TIM-4)gene in the immune response of oblique grouper  Epinephelus   coioides ,protein-coding regions of the oblique grouper was obtained using polymerase chain reaction (PCR),and the structural characteristics of its protein coding region were analyzed by bioinformatics.The tissue distribution of  TIM -4 gene were detected by real-time fluorescence quantitative PCR (qPCR).The expression profiles of  TIM -4 gene was investigated in spleen (GS) cells exposed to lipopolysaccharide (LPS) and polyinosinic acid (Poly I:C) stimulation.The recombinant plasmid  TIM -4-4T was constructed,and transformed into  Escherichia   coli  BL21 (DE3) for inducing prokaryotic protein expression.The results showed that the protein coding region (GenBank accession number:MZ465580) of  TIM -4 was 702 bp,encoding 234 amino acids protein containing three functional domains,a V-Type(IGV)domain,a BTK domain and a PRY domain.qPCR results showed that  TIM -4 was expressed in all examined 10 tissues of oblique grouper,with the maximal expression in stomach ( P <0 . 05),and the minimal in spleen.There was significantly lower expression of  TIM -4 in GS cells exposed to LPS stimulation,and significantly increase in GS cells exposed to Poly I:C stimulation ( P <0 . 05).The SDS-PAGE analysis revealed that the TIM-4 fusion protein had relative molecular weight of about 25 100.The optimal expression TIM-4 recombinant protein were inducing with 0.4 mmol/L IPTG at 37 ℃ for 6 h.The findings indicate that  TIM -4 may be involved in anti-infectious immunity of oblique grouper,which provides a theoretical basis for further study on the biological function of  TIM -4.
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Effects of low temperature stress on survival and expression of fatty acid metabolism-related genes SCD，FAD and ACC indifferent populations of Manila clam Ruditapes philippinarum
LI Ning, NIE Hongtao, LI Qiang, HUO Zhongming, YAN Xiwu
Journal of Dalian Ocean University    2022, 37 (4): 636-642.   DOI: 10.16535/j.cnki.dlhyxb.2021-243
Abstract （6243）   HTML （74）    PDF（pc） （2777KB）（3471）       Save
In order to probe into the effects of low temperature stress on the survival and fatty acid metabolism related genes of Manila clam  Ruditapes   philippinarum  zebra clam No.2, the survival differences and three genes related to fatty acid metabolisms including stearoyl coenzyme A desaturase ( SCD ), fatty acid desaturase ( FAD ) and acetyl coenzyme A carboxylase ( ACC ) in gill tissues were investigated in three populations including zebra clam No.2, Dalian in Liaoning Province and Beihai in Guangxi under low temperature stress of -1 (in a refrigerator)， 4 (in a recirculating tank)， and 12 ℃ (room temperature) by qRT-PCR. The results showed that after acute low temperature (-1, 4 ℃) stress for 9 d, the order of the survival rates of Manila clam populations was expressed as zebra clam No.2 > Dalian population in Liaoning > Beihai population in Guangxi. The expression of three fatty acid metabolism related genes in gill tissues was significantly affected by temperature ( P <0.05 ).With the temperature decreasing from 12 ℃ to -1 ℃, the expression level of the three genes were increased under 4 ℃ or -1 ℃ cold stress. Compared with 0 h, the expression levels of  SCD  and  FAD  genes in gill tissues of zebra clam No.2, Beihai population and Dalian population were significantly up-regulated from 12 to 72 h( P <0.05). The expression levels of  ACC  genes in the three populations were significantly up-regulated at different time, and the up-regulation range of the three genes in the gills of zebra clam No.2 was significantly higher than that in Beihai population and Dalian population ( P <0.05). The results demonstrated that the survival rate and low temperature tolerance of zebra clam No.2 at -1 ℃ were apparently higher than Dalian, Liaoning and Beihai, Guangxi. In addition, the high expression level of  SCD ,  FAD  and  ACC  genes of zebra clam No.2 might be related to its strong tolerance and high resistance to low temperature.
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Effects of dietary stickwater hydrolysate and plant essential oils on growth performance,immunity,digestive enzyme activity and TOR related gene expression of juvenile turbot Scophthalmus maximus
HAO Tiantian, XU Cong, WANG Jiying, LIU Caili, SHEN Yubo, LI Baoshan, SUN Yongzhi, HUANG Bingshan
Journal of Dalian Ocean University    2022, 37 (3): 441-449.   DOI: 10.16535/j.cnki.dlhyxb.2021-216
Abstract （5947）   HTML （141）    PDF（pc） （1217KB）（3467）       Save
The purpose of this experiment is to investigate the dietary stickwater hydrolysate (SWH) and plant essential oil on growth performance,immunity,digestive enzyme activity and TOR related gene expression of juvenile turbot  Scophthalmus   maximus  fed low fish meal diets.Juvenile turbot with initial body weight of 37.67 g were reared in a closed water circulating system and fed diets containing 50% fish meal (D1,positive control),and containing 30% fish meal (D2,negative control),and 30% fish meal diets supplemented with 1% SWH (D3),0.02% plant essential oil (D4),and 1% SWH+0.02% plant essential oil (D5) with triplication at water temperature of (16±1)℃ for 8 weeks.The results showed that there were significant increase in weight gain rate,specific growth rate and protein efficiency ratio and significant decrease in feed conversion ratio in the turbot in D3 group ( P <0.05).The maximal activities of serum lysozyme was observed in the turbot in D5 group ( P <0.05),significantly higher than that in the D2 group ( P <0.05),without significant difference from D1 group( P >0.05).The turbot in D5 group had the minimal C3 content,significantly lower than that in other 4 groups( P <0.05).The maximal intestinal trypsin activity was found in the turbot juveniles in D3 group ( P <0.05),significantly higher than that in the D2 group( P <0.05),without significant difference from D1 group ( P >0.05).The maximal  TOR  mRNA expression level was observed in liver of fish in D3 group,significantly higher than that in the D2 and D1 groups ( P <0.05),with an opposite trend in 4 EBP 1 and 4 EBP 2 mRNA expression levels ( P <0.05).There was significantly lower cumulative mortality of juveniles exposed to  Vibrio   anguillarum  infection at a dose of 7.94×10 5  CFU/g for 48 h in D4 and D5 groups than that in D2 group ( P <0.05).It can be concluded that supplementation of 1% SWH led to significantly enhance the growth performance,trypsin activity,liver TOR related gene expression level of turbot,and that 0.02% plant essential oil to significantly enhance the ability of fish to resist  V.anguillarum  infection.However,the combination of 1% SWH and plant essential oil does not result in good effect instead of low weight gain,the mechanism of their interaction needs further research.
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Characteristics and phylogenetic analysis of mitogenome in Atyidae
ZHU Longqiang, ZHU Zhihuang, ZHU Leiyu, WANG Dingquan, WANG Jianxin, LIN Qi
Journal of Dalian Ocean University    2022, 37 (3): 428-434.   DOI: 10.16535/j.cnki.dlhyxb.2021-177
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In this study,the complete mitogenome sequences of 24 species in Atyidae whose members are small in size,and difficult to make the morphological identification and classification were analyzed by using DnaSP 6,PhyML 3.1,MEGA 5.0,MrBayes 3.2.6 and BEAST 2.6.3 software.The results showed that the lengths of 24 mitogenomes were shown to be changed from 15 318 bp to 16 430 bp,with AT content varying from 62.20% to 70.19% and consistent gene composition and arrangement of mitogenome in 24 species.The analysis of differing sites of mitogenome revealed that the ratio of variable sites  nd 5 and  nd 4 genes were relatively high,with the maximal number of variable sites.The phylogenetic trees based on 13 PCGs (protein-coding genes) showed that the subfamily of Paratyinae was a monophyletic,and that Typhlatyinae and Caridellinae were non-monophyletic which were closely related to each other and clustered into a large branch,forming a sister group with Atyinae alone.Molecular clock analysis suggested that the most recent common ancestor of species in Paratyinae,Typhlatyinae and Caridellinae might occur from the middle Carboniferous to Permian time,while the most recent common ancestor of species in Atyinae might occur from the early Permian Jurassic time,and then further differentiated into the modern characteristic species in Atyidae.In conclusion, nd 5 and  nd 4 genes can be used as ideal molecular markers for species genetic analysis in species in Atyidae.The monophyleticity of Typhlatyinae and Caridellinae is still controversial,and the movement of geographic plates has an important impact on the species differentiation in Atyidae.
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Effects of alginate on biofilm formation of Pseudoalteromonas marina and larval settlement and metamorphosis of the mussel Mytilus coruscus
CHEN Huie, HE Kang, HE Chuhan, ZHU Youting, YANG Jinlong, LIANG Xiao
Journal of Dalian Ocean University    2022, 37 (4): 620-626.   DOI: 10.16535/j.cnki.dlhyxb.2021-260
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In order to research effects of alginate on biofilm formation of bacterium  Pseudoalteromonas   marina  and the roles of co-formed biofilm on larval settlement and metamorphosis of larval mussel  Mytilus   coruscus , the settlement and metamorphosis of the mussel larvae were investigated on biofilm of  P.marina  as a control, and the bacterial biofilm containing alginate solution at final concentration of 0.1, 0.5, 1.0, 1.5 and 2.0 mg/L. In addition, the optimal concentration of alginate was selected to compare and analyze the changes in density, thickness and extracellular products of the biofilm. The bacterial density on the biofilms was shown to be increased with the increase in the alginate concentration. There were significantly increase in thickness and extracellular polysaccharide contents of the biofilm at alginate concentration of 0.5 mg/L ( P <0.05), and finally increase in biofilm-inducing activities. The findings have positive significance for understanding of the induction mechanism of marine shellfish larval settlement and metamorphosis, and provide theoretical basis for the use of biofilms to improve the success rate of mussel seedlings. The appropriate concentration of alginate is 0.5 mg/L.
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Isolation,identification and virulence gene of atypical Aeromonas salmonicida from cultured turbot Scophthalmus maximus
WANG He, WANG Wenhao, HUANG Hua, LIU Xiaodan, ZHANG Liwen, YANG Tao, HU Lebin, LI Zhanjun
Journal of Dalian Ocean University    2022, 37 (4): 558-567.   DOI: 10.16535/j.cnki.dlhyxb.2021-254
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 In order to study the characteristics of atypical  Aeromonas   salmonicida , a dominant strain BHAS-1 was isolated and purified from the spleen of diseasing turbot individuals using conventional methods. The isolated strain BHAS-1 was identified by morphological observation, physiological and biochemical tests, and PCR specific amplification. The virulence gene and antibiotic sensitivity were also detected. The results showed that BHAS-1 strain was gram-negative cocci with β-hemolytic and no motility. The isolated stain BHAS-1 had similar physiological and biochemical properties to the atypical  A.salmonicida , but the strain did not produce indole and cannot utilize mannose. After 16S rRNA,  gyrB  and  vapA  sequence alignment analysis, BHAS-1 was identified as  A.salmonicida  subsp.  masoucida , and the virulence genotype was  aer + - zot + - dna + - ser + - lip + - sp + - alt + - act + - ast + - epa + - fla + - ahyB + , with LD 50  of 1.62×10 8  CFU/mL. The strain BHAS-1 was sensitive to 13 kinds of antibiotics such as enrofloxacin and tetracycline, resistant to 7 kinds of antibiotics such as puromyn and chloramphenicol, and moderately sensitive to ampicillin and streptomycin, with average resistance rate of 31.82%. In this study, we first reported the infection of  A.salmonicida  subsp.  masoucida  to cultured turbot in China. The molecular characteristics and drug sensitivity of the strain BHAS-1 were studied to provide reference for the prevention of furunculosis caused by atypical  A.salmonicida . The findings indicate that the application of vap gene can effectively identify different subspecies of  A.salmonicida .
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Regulatory mechanism of neuroendocrine system on immune response in molluscs:a review
WANG Lingling
Journal of Dalian Ocean University    2022, 37 (3): 363-375.   DOI: 10.16535/j.cnki.dlhyxb.2022-140
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The neuroendocrine system plays an important regulatory role in the metabolism,growth,development,reproduction and immunity response of animals.The reciprocal regulatory relationship between the neuroendocrine system and the immune system has been increasingly concerned in the fields of neurology,endocrinology,physiology,immunology and even medicine in recent years.Recently,the structure and function of the neuroendocrine system and its immunomodulatory mechanisms in invertebrates have attracted attention.Molluscs are eucoelomatous animals with abundant diversity,strong adaptability and wide distribution.It is found that molluscs have not yet differentiated into nerve and endocrine organs,but have evolved molecular mechanism similar to the neuroendocrine immune system in vertebrates,which can be a vital model to study the origin and evolution of invertebrate neuroendocrine system.This paper describes the molecular composition and activation mechanism of the molluscan neuroendocrine system and its regulatory effect on the immune response,proposes that the structure,function,regulation mechanism and evolutionary significance of molluscan neuroendocrine system remains significantly concern in the future research,aiming to provide reference for further understanding the evolution of neuroendocrine system and NEI-mediated environmental adaptation mechanism in molluscs.
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Effects of dissolved oxygen concentration on behavior, antioxidant enzyme activities and tissue structure of whelk Neptunea cumingii Crosse
ZHANG Qianhong, WANG Shaojun, TIAN Ying, WANG Luo, MAO Junxia, WANG Xubo, WANG Qingzhi, CHANG Yaqing, HAO Zhenlin
Journal of Dalian Ocean University    2022, 37 (4): 643-649.   DOI: 10.16535/j.cnki.dlhyxb.2021-309
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In order to explore physiological and ecological effects and to clarify the adaptation mechanism of whelk  Neptunea   cumingii  Crosse under different levels of dissolved oxygen (DO), the behavioral characteristics, histopathological damage in gill and olfactory tissue and antioxidant enzyme activity were investigated in the whelk with body weight of (85.3±9.3)g exposed to DO concentrations of 0, 2, 4, 6 and 8 (control) mg/L. The behavioral states of the whelk was divided into 4 types: normal state (less movement and long-term static adsorption), active state (short resting time, and long movement time), desorption state (foot edema), and death state (soft body part cannot be fully retracted into the shell). The whelk crawled actively and fed less when DO concentration was decreased, stopped feeding and death occurred at DO≤4 mg/L. Histological observation showed that the gap between gill filaments and the number of gill mucus cells were increased first and then decreased with the decrease in DO concentration. The top of gill filaments expanded, the respiratory epithelial cells fell off, and the surface cells sagged and formed folds, sometimes the gill filament was broken; the number of mucus cells was increased first and then decreased in osphradium with the decrease in DO concentration, without significant change in cell structure. The total antioxidant capacity (T-AOC) and activity of superoxide dismutase (SOD) in gill and hepatopancreas were shown to be decreased firstly, then increased and then decreased with the decrease in DO concentration, and the activity of CAT in gill to be increased first and then decreased. The activity of CAT in hepatopancreas was positively correlated with DO concentration. It was found that 6 to 8 mg/L of DO concentrations was the optimal or the whelk adapted to the hypoxic environment stress by changing behavior and related organizational structure, as well as regulating. The whelk was dead at DO≤4 mg/L for≥96 h.
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Spatio-temporal pattern and influencing factors of phytoplankton community structure in National Aquatic Germplasm Resources Reserve of common carp Cyprinus carpio and longsnout catfish Leiocassis longirostris in Huaihe River
WANG Dongwei, GONG Jiang, YANG Xiaoxi, ZHAO Wei, ZHOU Yanfeng
Journal of Dalian Ocean University    2022, 37 (4): 658-667.   DOI: 10.16535/j.cnki.dlhyxb.2021-311
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 In order to understand the spatio-temporal pattern of phytoplankton community structure and its relationship with environmental factors in common carp  Cyprinus   carpio  and longsnout catfish  Leiocassis   longirostris  National Aquatic Germplasm Resource Reserve in the Huaihe River (117°05′39″-117°15′31″E, 32°47′59″-33°00′03″N), phytoplankton and water quality were seasonally investigated in 18 sampling sites in the reserve in winter (January), spring (April), summer (August) and autumn (October) 2020.The results showed that 143 species of phytoplankton in 71 genera in 8 phyla, mainly Cyanophyta, Bacillariophyta and Chlorophyta, accounting for 85.31% in total, were identified in the reserve during the survey period. The dominant species of phytoplankton were mainly  Cyclotella   meneghiniana ,  Chroomonas   acuta  and other 13 species. The variation of Shannon-Wiener diversity index of phytoplankton in the reserve was ranged from 1.48 to 3.08, with significantly lower diversity index in spring and winter than that in summer and autumn ( P <0.05); Pielou evenness index was ranged from 0.46 to 0.96, without significant seasonal difference ( P >0.05). The range of Margalef richness index was varied from 0.48 to 2.58, significantly lower in winter than in the other three seasons, with the maximum in summer and the minimum in winter. The phytoplankton cell density in the reserve was ranged from 9.84×10 4  to 7.86×10 7  cells/L, and the variation range of biomass was varied from 0.02 to 29.07 mg/L. There was no significant difference in phytoplankton density and biomass in seasons, but significant difference in the waters of Guohe River, higher than that in the main stream of Huaihe River ( P <0.05). The main environmental factors affecting the spatio-temporal distribution of phytoplankton communities in the reserve were transparency (SD), total nitrogen (TN), water depth (WD) and permanganate index (COD Mn ) in spring, summer, autumn and winter, respectively. It was found that the watersheds in the reserve were all in a state of mild eutrophication, without significant difference in the phytoplankton density and biomass in time pattern, significantly higher in the Guohe River than that in the main stream of the Huaihe River. It is suggested that the corresponding environmental factors should be paid attention in different seasons to maintain the diversity of phytoplankton.
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Journal of Dalian Ocean University    2022, 37 (4): 707-718.  
Abstract （3741）      PDF（pc） （5782KB）（3411）       Save
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Construction of intelligent fish information sharing platform
LI Ran, YANG Yuting, ZHANG Zhiqiang, LIU Ying, HUANG Jianlong, LI Haomiao
Journal of Dalian Ocean University    2022, 37 (3): 497-504.   DOI: 10.16535/j.cnki.dlhyxb.2021-142
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In order to build an intelligent fish information sharing platform and to realize efficient and accurate retrieval of fish information,according to the characteristics of large amount of information and high concurrency of fish database,an intelligent fish information sharing platform is developed based on MongoDB.In the platform database 16 tables are designed,covering a variety of information such as fish name,ecology and morphology.The picture fish recognition is realized using the Java integrated framework of SpringBoot structure,and by integrating artificial intelligence fish recognition module,and convolution neural network.The management of loose data structure is realized by using the binaryjson of MongoDB,the mongotemplate service of spring is used for the database operation of MongoDB,and the network deployment is completed by using the cloud server platform provided by Alibaba cloud,as well as the combination of nginx reverse proxy and Tomcat server built in SpringBoot.The results showed that the database design realized the scientificity,traceability and practicability of fish information,reducing the coupling degree of the architecture and improving the maintainability of the program.The accuracy of fish identification by pictures was 92.67%.The system realizes the requirements of massive data storage and high concurrent access.The intelligent fish information sharing platform constructed in this study can provide a variety of retrieval entries based on compendium,region,morphology,fishing season,and presence or absence of scales.Users can query or identify the relevant information of the fish by uploading the text and picture of the fish.The detailed information of the fish can also be searched in the map according to the location of the fish,which meets the diversified retrieval needs.The platform information is open to realize the sharing of high-quality fish resources.The finding shows that the intelligent fish information sharing platform constructed in this study can effectively solve the problems of massive fish data storage and conveniently and effectively access the rich information.
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Cloning of X-linked inhibitor of apoptosis protein XIAP gene and its response to stimulation in sea urchin Strongylocentrotus intermedius challenged with pathogenic microorganisms
LIU Shengmei, SHANG Fengqin, CHEN Yadong, CHANG Yaqing, WANG Xiuli, QIU Xuemei, LIU Yang
Journal of Dalian Ocean University    2022, 37 (4): 627-635.   DOI: 10.16535/j.cnki.dlhyxb.2021-303
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 To investigate the role of X-linked inhibitor of apoptosis protein (XIAP) gene in the natural immune system of sea urchin  Strongylocentrotus   intermedius , the cDNA sequence of  XIAP  gene was cloned using rapid amplification of cDNA ends (RACE) technique, and distribution in various tissues including tube foot, peristomial membrane, intestine, and gonad and the expression of  XIAP  gene were analyzed in coelomocyte in the sea urchin exposed to stimulation including lipopolysaccharide (LPS), peptidoglycan (PGN), polyinosinic-polycytidylic acid (Poly I:C),  Vibrio   fortis  and whole glucan particles (WGP) by real-time quantitative PCR. The results showed that the length of  XIAP  cDNA was 4 894 bp, with 5′ UTR of 459 bp, the open reading frame of 2 331 bp and the 3′ UTR of 2 104 bp, encoding a total of 776 amino acids with three BIR structural domains and one RING finger structural domain, with a predicted relative molecular mass of 86 960 and an isoelectric point of 5.96, as acidic protein.  XIAP  was expressed in all tissues examined, significantly higher expression in the ovary than that in the tube foot, coelomocyte, peristomial membrane, intestine and testis ( P <0.05), after stimulation with PGN, LPS, Poly I:C, WGP and  V.fortis .  XIAP  gene expression was up-regulated in coelomocyte significantly ( P <0.05), with the maximal values 12 h after PGN, LPS and Poly I:C stimulation, and with the maximal values 24 h after WGP stimulation and 6 h after  V.   fortis  stimulation. The findings indicated that the clonally obtained  XIAP  gene was involved in the immune response of the sea urchin and played an important role in its natural immunity.
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Cloning and tissue expression of liver-expressed antimicrobial peptide Leap-2 in Ussuri cisco Coregonus ussuriensis exposed to bacterial infection
LIU Enhui, HUANG Tianqing, GU Wei, WANG Bingqian, ZHANG Yanping, LI Chunyu, XU Gefeng
Journal of Dalian Ocean University    2022, 37 (3): 420-427.   DOI: 10.16535/j.cnki.dlhyxb.2021-163
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To understand the molecular structure and expression characteristics of liver-expressed antimicrobial peptide  Leap -2 in Ussuri cisco  Coregonus   ussuriensis  Berg,the open reading frame (ORF) of the  Leap -2 gene was amplified from the liver tissue of Ussuri cisco with body weight of (30.0±1.2)g by PCR,and the Leap-2 amino acid sequence was analyze by bioinformatics software.The relative expression levels were detected in healthy tissues and tissues of Ussuri cisco intraperitoneally infected by  Aeromonas   hydrophila , Aeromonas   salmonicida  or  Staphylococcus   aureus  at a dose of 100 μL/ind.of 1×10 6  CFU/mL by fluorescence quantitative real-time PCR (qRT-PCR).The results showed that  Leap -2 gene (GenBank:MZ170050) had ORF region of 276 bp,which was cloned from the liver of Ussuri cisco by PCR.Leap-2 encoding 91 amino acids and its mature peptide region contained four highly conserved cysteine residues.Phylogenetic analysis showed that Ussuri cisco Leap-2 amino acid sequence was closely related to coho salmon  Oncorhynchus   kisutch ,with the homology of 96.7%.Tissue expression showed that Ussuri cisco  Leap -2 was the most abundant in the liver,followed by the intestine and kidney,with the minimal expression level in muscle.The up-regulated expression of  Leap -2 was observed in all detected Ussuri cisco exposed to  A.hydrophila , A.salmonicida ,and  S.aureus  infection.Western blot revealed that the expression of Leap-2 protein was up-regulated within 24 h,indicating that  Leap -2 was involved in the immune response of Ussuri cisco to bacteria.The finding may provide theoretical reference with understanding of the immune response of Ussuri cisco.
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Morphology and oxygen supply characteristics of gills in tuna: a review
FU Wenya, WEI Lu, ZHANG Pengfei, XIAO Juan, HUANG Hai, MA Zhenhua, GUO Zhiqiang
Journal of Dalian Ocean University    2022, 37 (5): 884-896.   DOI: 10.16535/j.cnki.dlhyxb.2021-258
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Tuna ( Thunnus  spp.) as pelagic fish with fast swimming are widely distributed throughout the temperate and tropical epipelagic waters of the world’s oceans. They require a large amount of oxygen supply to generate energy for fast swimming via gills as the oxygen supply organ. Focusing on the morphology and function characteristics of the gills during rapid swimming in tuna, this review descries the morphological structure of gills, gills surface area, oblique blood flow pattern and thin water blood barrier thickness structure and oxygen supply characteristics of related research. Further, to elucidate the structural adaptation (gill fusion, calcification, lamellar arrangement of gills) of tuna gills to high-speed water impact and the characteristics of gills countercurrent exchange. The morphological structure and problems in the field of oxygen supply for tuna gills are discussed and some progresses are suggested including the early tissue development of tuna gills, function of gills cells, mechanism of adaptive evolution of gills by bioinformatics and the relationship between tuna gills physiological processes and environmental factors to provide theoretical reference for further studied on the molecular mechanisms of tuna gills.
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Cloning and expression analysis of caspase-3 gene in development,immune and stress of variously colored abalone Haliotis diversicolor
LU Xiqin, ZHANG Lili, HUANG Shiyu, WANG Guodong, WANG Yilei, HE Simei
Journal of Dalian Ocean University    2022, 37 (3): 411-419.   DOI: 10.16535/j.cnki.dlhyxb.2021-141
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In order to explore the role of caspase apoptosis pathway in early development,immunity and stress of variously colored abalone  Haliotis   diversicolor ,a full-length cDNA sequence of  caspase -3 gene was cloned by rapid amplification of cDNA ends.qPCR was used to analyze the expression of  caspase -3 gene in various tissues,early developmental stages,infection with  Vibrio   parahaemolyticus ,thermal stress and hypoxia stress.The results showed that the full length of  caspase -3 cDNA was 1 192 bp,with encoding 252 amino acids.The encoding protein has highly conserved P20 and P10 active sites.There was a constitutive expression in detected tissues and in early developmental stages,with the highest expression in the hemolymph and in the fertilized egg and 2-cell stage ( P <0.05),respectively.The expression level of  caspase -3 in the hemolymph was significantly increased at 12 h after  V.parahaemolyticus  infection ( P <0.05).After thermal stress,the expression of  caspase -3 in hemolymph and hepatopancreas was significantly up-regulated at 24 h ( P <0.05).After hypoxia stress,the expression of  caspase -3 in gill tissue was significantly up-regulated at 96 h ( P <0.05),while the expression of  caspase -3 in hemolymph was significantly increased at 24 h ( P <0.05).The results suggest that  caspase -3 might play an important role in early development,immunity and stress of  H.diversicolor.
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Carbon storage assessment of sea grass Suaeda salsa community in Liaohe Estuary wetland based on HY-1C CZI data
LI Wei, WANG Wenshuo, ZHANG Xinyu, DUAN Yuanpeng, YANG Jianbo
Journal of Dalian Ocean University    2022, 37 (4): 574-583.   DOI: 10.16535/j.cnki.dlhyxb.2022-064
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 In order to use the HY-1C CZI data with higher spatial and temporal resolution to accurately assess the carbon storage and carbon sequestration efficiency of sea grass  Suaeda   salsa  in Liaohe Estuary wetland (121°23′-122°29′E， 40°39′-41°27′N), the spectral data and samples of  Suaeda   salsa  were collected in September of 2013 and 2018, and the biomass and carbon content were measured. The aboveground biomass inversion model was constructed based on transformed soil adjusted vegetation index (TSAVI) by regression analysis. The allometric growth model was used to estimate the belowground biomass, and then to estimate the carbon storage and carbon sequestration income of  Suaeda   salsa  in Liaohe Estuary wetland. The results showed that linear model was the best inversion model for aboveground biomass of  Suaeda   salsa  in Liaohe Estuary wetland. The coefficient of determination  R 2  of the model was 0.69 ( P <0.05), the values of root mean square error ( E ) and predictable skill (Skill) of the ground data verification set were 0.078 and 0.929, the values of relative root mean square error ( E r ) and Skill of the satellite data verification set were 9.55% and 0.987. It was estimated that the aboveground biomass of  Suaeda   salsa  community in Liaohe Estuary wetland in 2018, 2019 and 2020 were 1.69×10 6 , 1.13×10 6  and 3.26×10 6  kg, the belowground biomass of  Suaeda   salsa  community were 3.02×10 5 , 2.03×10 5  and 5.81×10 5  kg, the total biomass were 1.99×10 6 , 1.33×10 6  and 3.85×10 6  kg, and then the annual carbon storage of  Suaeda   salsa  community were 6.06×10 5 , 4.06×10 5  and 1.17×10 6  kg. The findings indicate that based on HY-1C CZI data to evaluate the carbon storage of  Suaeda   salsa  is feasible. The temporal and spatial distribution of carbon storage is consistent with its biomass distribution. The aboveground part of  Suaeda   salsa  is the main carbon storage part. The  Suaeda   salsa  in the Liaohe Estuary wetland has high ecological and economic benefits. This study has important reference value for the study of the blue carbon sink ecosystem in Liaohe Estuary wetland and the realization of carbon neutrality goal, and can provide technical and data support for the assessment of carbon storage in Liaohe Estuary wetland.
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Development and application of monoclonal antibody against IgM of silver prussian carp Carassius auratus gibelio
GUO Baoqin, WEI Chang, WANG Yinan, LI Qiang
Journal of Dalian Ocean University    2022, 37 (3): 450-456.   DOI: 10.16535/j.cnki.dlhyxb.2021-136
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In this study,immunoglobulin IgM from the serum of silver prussian carp  Carassius   auratus   gibelio  was isolated and purified by ammonium sulfate salting out method combined with protein A affinity chromatography.SDS-PAGE analysis showed that the purified IgM mainly contained two protein bands with relative molecular mass of 82 000 and 25 000,respectively.Balb/c mice were immunized with purified IgM,and five anti-crucian carp IgM monoclonal antibodies,named as 8G1,7C6,8C2,8C5 and 5C2,were obtained by hybridoma technique.Isotype analyzing indicated that 8G1,8C2,8C5 and 5C2 belonged to IgG1 isotype,and 7C6 belonged to IgG2b isotype.Western blot results showed that the five monodonal antibody (MAb) strains specifically recognized IgM light chain molecule with relative molecular mass of 25 000.The indirect immunofluorescence showed that all the five monoclonal antibodies recognized the sIgM cell of silver prussian carp.In addition,the indirect ELISA method for detection of the anti-CyHV-2 serum antibody in sliver carp was established using the anti-IgM monoclonal antibody as primary antibody and AP-labeled sheep anti-mouse Ig as the secondary antibody.The results of condition optimization indicated that the candidate 8C5 is the optimal anti-IgM monoclonal antibody,and 1∶20 dilution of fish serum is the optimal concentration for ELISA detection.Results of this study provides an effective tool for future research on immune response of silver prussian carp and an effective method for the evaluation of immune effect of vaccination.
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Distribution and relationship with influential factors of dominant zooplankton species in the Northwest Pacific Ocean in spring and winter
FU Feiyu, HAN Peiwu, FANG Zhou, LIU Bilin
Journal of Dalian Ocean University    2022, 37 (3): 489-496.   DOI: 10.16535/j.cnki.dlhyxb.2021-130
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In order to explore horizontal distribution characteristics of dominant zooplankton species in the Northwest Pacific Ocean,the dominant zooplankton was surveyed in zooplankton samples collected at 58 stations in the Northwest Pacific Ocean (146°-156°E,28°-35°N) in spring (March) 2019 and winter (December)2020.The relationship between dominant species and influencing factors of zooplankton were evaluated by Pearson correlation analysis and redundancy analysis.The results showed that 602 species (including unknown species) were identified during the two voyages,with more number of species in spring (456) than that in winter (386).Eleven dominant species were observed,including  Pleuromamma   gracilis , Nannocalanus   minor , Neocalanus   gracilis  and  Sagitta   hexsaptera  in the two seasons.Pearson correlation analysis revealed that latitude had a great influence on spring when there was significant or very significant negative correlation between latitude and  Doliolum   nationalis  ( P <0.05 or  P <0.01). Mesocalanus   tenuicornis  and  Eudoxoides   spiralis  were negatively correlated with latitude ( P <0.05),and  Calanus   jashnovi  was positively correlated with latitude ( P <0.05). Lucicutia   flavicornis  and  S.hexsaptera  were positively correlated with salinity in the winter survey area.Redundancy analysis (RDA) was consistent results with Pearson correlation analysis,with the latitude as main factor affecting the abundance of dominant species in spring ( P <0.01),and salinity in winter ( P <0.05).The results showed that the horizontal distribution of dominant species in winter and spring in the Northwest Pacific Ocean was controlled by different factors,which was the result of the comprehensive influence of many factors.The finding indicated that distribution of dominant species in spring was deeply affected by latitude,while salinity was the key factor in winter.
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Differences in intestinal microbial community between Dolly varden trout Salvelinus malma and lenok Brachymystax lenok with different ages
LIU Peng, LI Lingxue, ZHENG Wei, DU Xiaoyan, YAN Chunmei, LIU Huiji, GAO Chunshan
Journal of Dalian Ocean University    2022, 37 (4): 568-573.   DOI: 10.16535/j.cnki.dlhyxb.2021-193
Abstract （5438）   HTML （7）    PDF（pc） （3868KB）（3340）       Save
In order to explore the intestinal microorganisms of Dolly varden trout  Salvelinus   malma  (1 +  and 2 +  years old) and lenok  Brachymystax   lenok  (4 +  years old), MiSeq 16S rRNA high-throughput sequencing was used to sequence and analyze the microbes in intestinal contents of healthy Dolly varden trout and lenok in same aquaculture environment. The results showed that there were 3 896 OTUs in the intestinal contents of the Dolly varden trout, and 1 675 OTUs in the intestinal contents of lenok,showing that the dominant microflora was rich in the contents of Dolly varden trout and lenok. There was higher diversity in the 1 +  year old Dolly varden trout groups (H1) than that in the 2 +  years old groups (H2), indicating that the intestinal microorganisms were planted in the host intestines with the growth, with the dominant flora  Mycoplasma  in the Dolly varden trout, and the dominant flora  Bacillus  in the lenok. PCA analysis revealed that there was the similar intestinal microbial community structure in 1 +  and 2 +  years old Dolly varden trout groups, and different from the 4 +  years old lenok (X4). The findings indicated that the fish of same species with different ages had similar intestinal microbial composition under the same culture environment, and the quite different microbial composition was observed in the two salmon species in the same family and different genera, though with the similar microbial diversity.
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Functional divergence of different ep300/crebbp gene family members in rainbow trout Oncorhynchus mykiss
WANG Xianzong, LIU Qing, LI Shu
Journal of Dalian Ocean University    2022, 37 (3): 376-385.   DOI: 10.16535/j.cnki.dlhyxb.2021-169
Abstract （5468）   HTML （1）    PDF（pc） （24227KB）（3339）       Save
To probe into copy number variation,functional divergence and correlation with hypoxia response of  ep 300 /crebbp  gene family in rainbow trout  Oncorhynchus   mykiss ,TBLASTN and BLASTP search and reconstruction of phylogenetic tree were firstly conducted,based on which the members and their orthologs in zebrafish  Danio   rerio  were determined.Tissue expression profiles and co-expressed genes of these copies were analyzed based on original transcriptome sequencing data,and GO enrichment analyses of these co-expressed genes were also conducted.The degree of variation among proteins encoded by these copies was compared by CDD search,multiple sequence alignment and three-dimensional structural modeling.The results showed that there were eight  ep 300 /crebbp  members in rainbow trout ( ep 300 aa - ep 300 bb , crebbpaa - crebbpbb ),which were all formed by whole genome duplication events and could be divided into four ohnolog pairs corresponding to the four members in  D.rerio .There were significant differences in tissue expression profiles,and primary and tertiary structures of encoded proteins between different ohnolog pairs,much smaller differences between two copies within an ohnolog pair.These eight members had different numbers of unique co-expressed genes,and the number of GO terms enriched by these gene collections related to binding,modification,stress/stimulus-response and hypoxia tolerance were also varied greatly. ep 300 ba  and  ep 300 bb  belong to an ohnolog pair,and yet their encoded proteins were varied in 35 positions in conserved domains,with the far greater number than other ohnolog pairs.Furthermore,only co-expressed genes of  ep 300 bb  enriched GO terms related to stress/stimulus-response and hypoxia tolerance.The findings suggest that  ep 300 bb  can serve as an important candidate gene for genetic improvement of rainbow trout due to its great evolution rate and stronger correlation with hypoxia tolerance than other  ep 300 /crebbp  members.
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Recognition of nested named entities in aquature medicine based on multi-kernel convolution (BERT+Multi-CNN+CRF)
LIU Jusheng, YU Hong, YANG Huining, SHAO Liming, SONG Qishu, LI Guangyu, ZHANG Sijia, SUN Hua
Journal of Dalian Ocean University    2022, 37 (3): 524-530.   DOI: 10.16535/j.cnki.dlhyxb.2022-047
Abstract （5210）   HTML （59）    PDF（pc） （3220KB）（3330）       Save
To address the problem of low accuracy of nested named entity recognition in named entity recognition in aquature medicine,a named entity recognition method is proposed based on multi-core convolutional neural networks (BERT+Multi-CNN+CRF).Multi-core convolutional neural networks are used to extract nested entity features,while BERT (bidirectional encoder representations from transformers) is used to pre-train the input corpus to obtain nested entity input feature matrix and enhance the position vector of nested entity.Then the extracted feature matrix is fused with the input feature matrix to enhance the feature representation of nested entities.Comparison experiments of different models verified the recognition effectiveness of the proposed model,with the accuracy of 88.04%,recall of 88.92% and F1 values of 88.48% in the proposed BERT+Multi-CNN+CRF model in the nested NER task in aquature medicine.Compared with the BERT+BiLSTM+ATT+CRF model which has good recognition ability,the accuracy,recall,and F1 value of the proposed model are improved by 2.25%,3.23%,and 2.74% respectively.The finding shows that the model proposed in this paper can effectively solve the problem of low accuracy of NER in aquaculture medicine,and is an effective method for nested NER recognition in aquatic medicine.
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Prokaryotic expression and acetylation identification of phosphomethylpyrimidine synthase (ThiC) in Vibrio alginolyticus
WANG Xudong, FAN Chenlong, DING Yu
Journal of Dalian Ocean University    2022, 37 (3): 435-440.   DOI: 10.16535/j.cnki.dlhyxb.2021-148
Abstract （5925）   HTML （161）    PDF（pc） （3977KB）（3302）       Save
To understand whether there is acetylation of phosphomethylpyrimidine synthase (ThiC) in pathogenic  Vibrio   alginolyticus ,the target gene  ThiC  was amplified via PCR basing on the genome of  V.alginolyticus  HY 9901.The prokaryotic expression vector pET-28a- ThiC  were constructed,which was successfully expressed in  Escherichia   coli  BL21 by induction,and its expression conditions were optimized.Then,the protein expression and acetylation level were analyzed by SDS-PAGE and Western blot.The results showed that the  ThiC  gene had length of 1 941 bp,and that the recombinant  Escherichia   coli  BL21 was expressed as the recombinant protein(72 500).The optimal expression of recombinant protein was under conditions of 0.1 mmol/L IPTG for 3 hours at 37 ℃.Western blot results revealed that ThiC was acetylated protein,and there was no deacetylation in vitro.In conclusion,the recombinant expression strain for  V.alginolyticus  ThiC was successfully constructed,ThiC was acetylated protein,and there was no deacetylation in vitro.The findings lay the foundation for studying the post-translational regulation mechanism of vitamin nutrition of  V.alginolyticus .
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Prediction of sea surface temperature based on ConvGRU deep learning network model
ZHANG Xuewei, HAN Zhen
Journal of Dalian Ocean University    2022, 37 (3): 531-.   DOI: 10.16535/j.cnki.dlhyxb.2021-137
Abstract （5461）   HTML （6）    PDF（pc） （6790KB）（3297）       Save
In order to solve the problems of insufficient combination of temporality and spatiality of traditional time series network models and insufficient ability of batch processing large data of marine environmental elements,sea surface temperature (SST) prediction and analysis were carried out using the SST grid data from 1999 to 2019 in part of the Northwest Pacific Ocean (OISST Products) by ConvGRU deep learning neural network model,that is,combination of the extended algorithm of recurrent neural network (RNN) and convolutional neural network (CNN),and by sample generator to process data,which can effectively deal with the batch problem of time series remote sensing data.The results showed that the ConvGRU model training set had root mean square error of 0.044 9 ℃ and accuracy of 99.69%,and that the validation set had root mean square error of 0.045 2 ℃ and accuracy of 99.64%.Finally,the model established in this paper was used to predict the sea surface temperature data in 2020,with the root mean square error of 0.047 8 ℃ and accuracy of 99.60% in the test set.The average mean absolute deviation of SST prediction was shown to be about 0.379 3 ℃,and the average prediction accuracy to be 97.31%.The findings indicate that the ConvGRU neural network model can predict the trend of sea surface temperature well,which provides a feasible method for SST neural network prediction model.
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Preparation and characterization of thermoresponsive starch ether/sodium alginate composite hydrogels
PANG Zijian, XU Yuan, JIANG Shuo, SUN Meng, JIANG Qian, SUN Wei, ZHA Xiaohan, SHANG Yue, DAI Mingyun, LIU Ying, TIAN Ye
Journal of Dalian Ocean University    2022, 37 (3): 505-512.   DOI: 10.16535/j.cnki.dlhyxb.2021-176
Abstract （5767）   HTML （4）    PDF（pc） （3937KB）（3290）       Save
A thermoresponsive hydrogel composed of 2-hydroxy-3-isopropoxypropyl starch (HIPS) and sodium alginate (SA) was synthesized via cross-linking with hybrid crosslinkers including ethylene glycol diglycidyl ether (EDGE) and calcium chloride (CaCl 2 ).Attenuated total reflection infrared spectroscopy (FT-IR) was used to confirm the occurrence of crosslinking.The HIPS/SA composite hydrogels were investigated for their porous architecture and thermoresponsive property using scanning electron microscopy (SEM).The reaction temperature and reaction time significantly affected the swelling rate of HIPS/SA composite hydrogels ( P <0.01).The optimum HIPS/SA hydrogel synthesis was observed under conditions of HIPS concentration of 6.5% (2 g),SA concentration of 4.5% (2 g),NaOH of 300 μL,EDGE of 300 μL,reaction temperature of 55 ℃ and reaction period of 3 h,with the hydrogel swelling ratio of 44.20 under above conditions.It was found that the HIPS/SA hydrogel possessed desirable thermoresponsive property,and the swelling rate (SR) of hydrogel was sharply decreased when the temperature was higher than volume phase transition temperature (VPTT) 34.9 ℃.The HIPS/SA hydrogel with thermoresponsive properties synthesised in this study can provide a new method for the application in the fields of drug delivery and separation engineering.
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Effects of flavor protease on quality of marinated mandarin fish at ice temperature
XU Ming, WAN Jinqing, HOU Boyu, WANG Youjun, SUN Xiaolin, TONG Nian
Journal of Dalian Ocean University    2022, 37 (3): 513-523.   DOI: 10.16535/j.cnki.dlhyxb.2021-173
Abstract （5508）   HTML （1）    PDF（pc） （1717KB）（3285）       Save
In order to explore the effect of flavor protease on physical and chemical properties,taste and smell of the marinated mandarin fish  Siniperca   chuatsi  under ice temperature (-2 ℃) conditions,changes in composition,sensory,amino acid nitrogen,trichloroacetic acid (TCA)-soluble peptides,free amino acids,flavor nucleotides,and volatile substances were determined in salted(3% of NaCl) mandarin fish steaks supplemented with flavor protease at a dose of 25,50 and 100 U/g and disposed in a sealed box at ice temperature (-2 ℃) for 10 d.The results showed that addition of flavor protease at ice temperature led to significant effect on the taste and flavor of pickled mandarin fish ( P <0.05),the higher the dose,the greater the effect,with the too high concentration of flavor protease resulting in flesh hardness and loss of taste.It was found that the optimal concentration of flavor protease was of 50 U/g,and under the optimal concentration the best fish quality index,and TCA-soluble peptide of 347.22 mg/100 g,1.5 times as those in the control group,were observed in the 50 U/g group.The maximal umami amino acid content of 60.78 mg/100 g was found on the 6th day,and the maximal inosine nucleotide (IMP) content of 133.83 mg/100 g on the 6th day,1.3 times as that in the control group,and the maximal MSG equivalent content of 7.06 g MSG/100 g on the 6th day,2.8 times as that in the control group.At the same time,there were more abundant volatile substances in the fish meat in the 50 U/g group,with total amount of 2 625.11 μg/100 g,2.7 times as those in the control group.The findings indicated that addition of flavor protease led to significantly improve the quality and flavor of the marinated mandarin fish under the best conditions of 50 U/g enzyme for 6 days,which provided better technology and new ideas for traditional pickled mandarin fish and theoretical reference for the ice temperature research of fishery products.
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Extraction, purification and pharmacological activity of Polysaccharides from sea weed Sargassum pallidum
KONG L, ZHU Yuexia, ZHOU Bingxue, LIU Zhi, XUE Wangqian, SHI Jinfeng, ZHENG Jiaqin, LI Jiaojiao, JI Jing, QIN Kunming, DONG Zibo, SHEN Jinyang
Journal of Dalian Ocean University    2022, 37 (5): 897-.   DOI: 10.16535/j.cnki.dlhyxb.2021-184
Abstract （4804）   HTML （4）    PDF（pc） （1918KB）（3285）       Save
Sargassum   pallidum  as a typical representative of brown seaweed is widely distributed in the coastal areas of China including the Yellow Sea and the Bohai Sea, and is one of the earliest Marine Traditional Chinese medicines. Brown seaweed  S.pallidum  has several effects including resolving phlegm, softening and firmness, eliminating galls and nodules, promoting diuresis and detumescence, and has been used in the treatment of phlegm edema, gall, beriberi, adverse urination and other diseases. In recent years, there have been a lot of research on the methods of extraction and purification of polysaccharides from  S.pallidum , as well as the biological activities of antioxidant, hypoglycemic, antitumor, antibacterial, hypolipidemic and immunomodulatory. The purified polysaccharide of  S.pallidum  was primarily composed of fucose, galactose, xylose, glucose, mannose and arabinose. In this paper, the extraction, separation and purification of polysaccharides from  S.pallidum , as well as pharmacological activities are reviewed. Given of the problems existing in the research, suggestions for the future development of the optimization of the method of extraction and purification of the polysaccharide, the research and development of the polysaccharide preparation, and in-depth research on the fine structure of the polysaccharide are put forward, so as to make some contributions to further development and utilization of  S.pallidum .
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Artificial propagation,embryonic and early larval development of Schizothorax eurystomus
XIA Zihui, ZHAO He, QIU Meng, NIE Zhulan, CHEN Kexun, LI Xuetao, QIN Xiaofang, WEI Jie
Journal of Dalian Ocean University    2022, 37 (3): 471-481.   DOI: 10.16535/j.cnki.dlhyxb.2021-132
Abstract （5621）   HTML （140）    PDF（pc） （6524KB）（3271）       Save
Broodstock  Schizothorax   eurystomus  collected from Kizil River Basin in Xinjiang Uygur Autonomous Region was induced to spawn by hormone injection of pituitary gland (PG),human chorionic gonadotropin (HCG) and luteinizing hormone-releasing hormone analogs (LHRH-A 2 )at various doses and artificial hatchery methods at water temperature from 10 ℃ to 20 ℃ in a 1 ℃ interval in 2019 to accomplish farming and stock enhancement of  S.eurystomus .The results showed that success rate and hatching rate were significantly varied with the doses of hormones and water temperature,with the optimal hormone combinations of PG at 10.27 mg/kg,HCG at 300.00 IU/kg and LHRH-A 2  of 19.41 μg/kg.In April and May 2019,the fertilized eggs of  S.eurystomus  were obtained by artificial propagation,and the development of embryo and larva was observed under artificial incubation conditions,with the optimum incubation water temperature of 16.0-17.0 ℃. S.eurystomus  had egg diameters of (2.50±0.24)mm,and reached (3.43±0.11)mm after water swelling;under the condition of water temperature (16.19±0.08)℃,the fertilized eggs were hatched in 145 h and 48 min,with the total accumulated temperature of 2 364.80 ℃·h.The embryonic development process experienced seven stages:embryo disc,cleavage,blastula,gastrula,neuroembryo,organogenesis and hatching.The newly hatched larvae had total length of (8.03±0.15) mm and the yolk sac total length of (20.17±0.96)mm,with the completely exhausted yolk sac at (17.88±0.44)℃ for 480 h.The findings further enrich the early life history data of  S.eurystomus  and then provide essential data including water temperature regulation,and choice of hormone and dose for resources protection,and large scale artificial propagation of  S.eurystomus.
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